Project Summary/Abstract The Bioanalytics Core laboratory within the University of Louisville Diabetes and Obesity Center (DOC) as well as other laboratories on campus operate mass spectrometers dedicated to meeting ever increasing and complex needs of NIH-funded investigators. The needs of our investigators far exceed current capabilities, with major limitations comprising a lack of ability for untargeted adductomic and metabolomic screening, lipidomics, and stable isotope metabolomics. These major applications are central to several NIH-funded Major and Minor Users with interests in cardiovascular and metabolic disease, environmental toxicology, exercise science, and immunology. We propose to add a Bruker timsTOF LC-MS system, which would significantly enhance our breadth of analyte coverage and open the possibility for biomarker discovery and identification of unknown endogenous and exogenous metabolites in biological samples. This will be important to Major Users engaged in biomarker discovery and the identification of conjugates of reactive species. In addition, the system would expand our coverage to hundreds of lipids within multiple lipid classes and would empower stable isotope metabolomic capabilities beyond central carbon metabolism. Features of the Bruker timsTOF LC-MS system, such as the high scan speed and ultrahigh resolution in the MS and MS/MS modes and 5 orders of magnitude dynamic range enables coverage of thousands of analytes in complex biological matrices while maintaining high throughput analyses. The High Resolution Ion Mobility further adds another dimension for separating ions, allowing for separation of isomeric and isobaric species ? a feature of special use in adductomics, metabolomics and lipidomics applications. This application describes the utility of the instrument to six Major Users, who are funded by large NIH multicenter grants and R01s, and several Minor Users. Proposed work with instrument includes detailed training and education in mass spectrometry and method development, which will further advance research in the Center and maximize the utility of the instrument. The DOC has the infrastructure to maintain and support the new equipment backed by the support of the University.